D-Amino acid oxidase knockdown in the mouse cerebellum reduces NR2A mRNA
| Autor: | Wood Mja., N Nikiforova, Paul Harrison, L Chen, Burnet Pwj., Patrick N. Anderson |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
D-Amino-Acid Oxidase
Male Genetic Vectors Green Fluorescent Proteins D-amino acid oxidase Biology Receptors N-Methyl-D-Aspartate Cell Line Serine Small hairpin RNA Mice Cellular and Molecular Neuroscience RNA interference Cerebellum Gene expression Animals RNA Messenger Molecular Biology Gene knockdown Oxidase test Messenger RNA Mental Disorders Lentivirus Cell Biology Molecular biology Rats Mice Inbred C57BL Protein Subunits nervous system Gene Knockdown Techniques RNA Interference |
| DOI: | 10.1016/j.mcn.2010.08.018 |
| Popis: | Virus mediated RNA-interference (RNAi) is a powerful approach to study genes in vivo. Here we report a method using lentivirus-delivered RNAi to knockdown the glial enzyme, d-amino acid oxidase (DAO), in the mouse cerebellum. After initial characterisation in vitro, we achieved a 40-50% reduction of DAO mRNA in the cerebellum 7 and 28. days after a single injection of lentivirus encoding a DAO-specific, short-hairpin RNA. Injections also decreased DAO immunoreactivity (-33%). The major substrate for DAO is d-serine, an N-methyl-d-aspartate receptor (NMDAR) co-agonist. Thus, we also measured whether DAO knockdown impacted on d-serine, or expression of NMDAR subunits, and found that DAO RNAi led to increased cerebellar d-serine levels (+. 77%), and decreased NMDAR subunit NR2A mRNA (-22%), but did not affect NR1 or NR2C mRNAs. These data demonstrate the utility of lentiviruses to deliver RNAi to glial cells within the cerebellum, and confirm the role of DAO in d-serine metabolism. They also provide a tool to investigate DAO, an enzyme currently of considerable interest in the pathophysiology and therapy of schizophrenia. © 2010 Elsevier Inc. |
| Databáze: | OpenAIRE |
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